IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ zhang-2000-1 The coiled-coil domain of Stat3 is essential for its SH2 domain-mediated receptor binding and subsequent activation induced by epidermal growth factor and interleukin-6. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Zhang T., Kee WH., Seow KT., Fung W., Cao X. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease Cancer - Interactions investigated in the context of cancer COS-1 cells were transfected with the Stat3 plasmids and lysed after 48 h. The biotinylated peptides derived from gp130 were incubated with streptavidin-Sepharose. The beads were washed and then incubated with aliquots of lysates. The complexes were washed, fractionated on SDS-PAGE, and immunoblotted with anti-Stat3 antibody. The blot was then stripped and reprobed with anti-SHP-2 antibody. mcbcaoxm@imcb.nus.edu.sg Mol. Cell. Biol. (0270-7306) 2000 zhang-2000-2 The coiled-coil domain of Stat3 is essential for its SH2 domain-mediated receptor binding and subsequent activation induced by epidermal growth factor and interleukin-6. in vitro In vitro enzymatic study enzymatic study predetermined predetermined participant predetermined Zhang T., Kee WH., Seow KT., Fung W., Cao X. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease Cancer - Interactions investigated in the context of cancer COS-1 cells were transfected with the Stat3 plasmids and lysed after 48 h. The biotinylated peptides derived from gp130 were incubated with streptavidin-Sepharose. The beads were washed and then incubated with aliquots of lysates. The complexes were washed, fractionated on SDS-PAGE, and immunoblotted with anti-Stat3 antibody. The blot was then stripped and reprobed with anti-SHP-2 antibody. mcbcaoxm@imcb.nus.edu.sg Mol. Cell. Biol. (0270-7306) 2000 zhang-2000-3 The coiled-coil domain of Stat3 is essential for its SH2 domain-mediated receptor binding and subsequent activation induced by epidermal growth factor and interleukin-6. in vitro In vitro pull down pull down antibody detection identification by antibody antibody detection Zhang T., Kee WH., Seow KT., Fung W., Cao X. IBD - Inflammatory bowel disease Crohn's disease - Interactions of proteins identified as having a link to Crohn's disease Cancer - Interactions investigated in the context of cancer COS-1 cells were transfected with the Stat3 plasmids and lysed after 48 h. The biotinylated peptides derived from gp130 were incubated with streptavidin-Sepharose. The beads were washed and then incubated with aliquots of lysates. The complexes were washed, fractionated on SDS-PAGE, and immunoblotted with anti-Stat3 antibody. The blot was then stripped and reprobed with anti-SHP-2 antibody. mcbcaoxm@imcb.nus.edu.sg Mol. Cell. Biol. (0270-7306) 2000 il6rb_human Interleukin-6 receptor subunit beta IL6ST Interleukin-6 signal transducer Membrane glycoprotein 130 CDw130 Oncostatin-M receptor subunit alpha protein protein human Homo sapiens Human MLTLQTWLVQALFIFLTTESTGELLDPCGYISPESPVVQLHSNFTAVCVLKEKCMDYFHVNANYIVWKTNHFTIPKEQYTIINRTASSVTFTDIASLNIQLTCNILTFGQLEQNVYGITIISGLPPEKPKNLSCIVNEGKKMRCEWDGGRETHLETNFTLKSEWATHKFADCKAKRDTPTSCTVDYSTVYFVNIEVWVEAENALGKVTSDHINFDPVYKVKPNPPHNLSVINSEELSSILKLTWTNPSIKSVIILKYNIQYRTKDASTWSQIPPEDTASTRSSFTVQDLKPFTEYVFRIRCMKEDGKGYWSDWSEEASGITYEDRPSKAPSFWYKIDPSHTQGYRTVQLVWKTLPPFEANGKILDYEVTLTRWKSHLQNYTVNATKLTVNLTNDRYLATLTVRNLVGKSDAAVLTIPACDFQATHPVMDLKAFPKDNMLWVEWTTPRESVKKYILEWCVLSDKAPCITDWQQEDGTVHRTYLRGNLAESKCYLITVTPVYADGPGSPESIKAYLKQAPPSKGPTVRTKKVGKNEAVLEWDQLPVDVQNGFIRNYTIFYRTIIGNETAVNVDSSHTEYTLSSLTSDTLYMVRMAAYTDEGGKDGPEFTFTTPKFAQGEIEAIVVPVCLAFLLTTLLGVLFCFNKRDLIKKHIWPNVPDPSKSHIAQWSPHTPPRHNFNSKDQMYSDGNFTDVSVVEIEANDKKPFPEDLKSLDLFKKEKINTEGHSSGIGGSSCMSSSRPSISSSDENESSQNTSSTVQYSTVVHSGYRHQVPSVQVFSRSESTQPLLDSEERPEDLQLVDHVDGGDGILPRQQYFKQNCSQHESSPDISHFERSKQVSSVNEEDFVRLKQQISDHISQSCGSGQMKMFQEVSAADAFGPGTEGQVERFETVGMEAATDEGMPKSYLPQTVRQGGYMPQ stat3_mouse Signal transducer and activator of transcription 3 Acute-phase response factor Stat3 Aprf protein protein mouse Mus musculus Mouse MAQWNQLQQLDTRYLEQLHQLYSDSFPMELRQFLAPWIESQDWAYAASKESHATLVFHNLLGEIDQQYSRFLQESNVLYQHNLRRIKQFLQSRYLEKPMEIARIVARCLWEESRLLQTAATAAQQGGQANHPTAAVVTEKQQMLEQHLQDVRKRVQDLEQKMKVVENLQDDFDFNYKTLKSQGDMQDLNGNNQSVTRQKMQQLEQMLTALDQMRRSIVSELAGLLSAMEYVQKTLTDEELADWKRRQQIACIGGPPNICLDRLENWITSLAESQLQTRQQIKKLEELQQKVSYKGDPIVQHRPMLEERIVELFRNLMKSAFVVERQPCMPMHPDRPLVIKTGVQFTTKVRLLVKFPELNYQLKIKVCIDKDSGDVAALRGSRKFNILGTNTKVMNMEESNNGSLSAEFKHLTLREQRCGNGGRANCDASLIVTEELHLITFETEVYHQGLKIDLETHSLPVVVISNICQMPNAWASILWYNMLTNNPKNVNFFTKPPIGTWDQVAEVLSWQFSSTTKRGLSIEQLTTLAEKLLGPGVNYSGCQITWAKFCKENMAGKGFSFWVWLDNIIDLVKKYILALWNEGYIMGFISKERERAILSTKPPGTFLLRFSESSKEGGVTFTWVEKDISGKTQIQSVEPYTKQQLNNMSFAEIIMGYKIMDATNILVSPLVYLYPDIPKEEAFGKYCRPESQEHPEADPGSAAPYLKTKFICVTPTTCSNTIDLPMSPRTLDSLMQFGNNGEGAEPSAGGQFESLTFDMDLTSECATSPM ptpn11-cerae-protein PTPN11 (SHP-2) African Green Monkey protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey An amino acid sequence which is not represented in UniProt Similar to Q06124 src_human Proto-oncogene tyrosine-protein kinase Src pp60c-src SRC SRC1 Proto-oncogene c-Src protein protein human Homo sapiens Human MGSNKSKPKDASQRRRSLEPAENVHGAGGGAFPASQTPSKPASADGHRGPSAAFAPAAAEPKLFGGFNSSDTVTSPQRAGPLAGGVTTFVALYDYESRTETDLSFKKGERLQIVNNTEGDWWLAHSLSTGQTGYIPSNYVAPSDSIQAEEWYFGKITRRESERLLLNAENPRGTFLVRESETTKGAYCLSVSDFDNAKGLNVKHYKIRKLDSGGFYITSRTQFNSLQQLVAYYSKHADGLCHRLTTVCPTSKPQTQGLAKDAWEIPRESLRLEVKLGQGCFGEVWMGTWNGTTRVAIKTLKPGTMSPEAFLQEAQVMKKLRHEKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKGETGKYLRLPQLVDMAAQIASGMAYVERMNYVHRDLRAANILVGENLVCKVADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELTTKGRVPYPGMVNREVLDQVERGYRMPCPPECPESLHDLMCQCWRKEPEERPTFEYLQAFLEDYFTSTEPQYQPGENL il6st-stat3 pull down of STAT3 by gp130 peptide pY3 567910 N/A 567913 unspecified role unspecified role bait bait required phosphotyr-767 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain false region gp130 pY3 peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 567914 unspecified role unspecified role prey prey c-terminal ST3-DN1H required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false P42227:p.Asp170Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P42227:p.Lys177Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 physical association physical association false false false Fig. 4b, Fig. 5b Phosphorylation of the tyrosine-767 in the gp130 peptide was required for the interaction. Binding of ST3-DN1H (lacking the first helix in the coiled-coil domain) was less than the full length protein and the deletion mutants ST3-DN2H (260-770), ST3-DN3H (296-770) and ST3-DN4H (320-770) did not bind at all. il6st-ptpn1 pull down of SHP-2 by gp130 peptide pY2 567910 N/A 567913 unspecified role unspecified role bait bait region gp130 pY2 peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 567915 unspecified role unspecified role prey prey chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 physical association physical association false false false Fig. 4c Phosphorylation of the tyrosine-759 in the gp130 peptide was required for the interaction. src-stat3 567911 N/A 567916 enzyme enzyme unspecified role unspecified role spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. N/A 567914 enzyme target enzyme target substrate substrate unspecified role unspecified role c-terminal ST3-DN1H sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction false false false Fig. 6a Stat3 is phosphorylated by Src on Tyr-705. The authors do not specify the species of origin of Src. However, the website of Upstate Biotechnology from where the enzyme was purchased shows an active human Src in the catalogue. il6st-stat3-1 pull down of STAT3 by gp130 peptide pY3 567912 N/A 567913 unspecified role unspecified role bait bait region gp130 pY3 peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false required phosphotyr-767 optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 567914 unspecified role unspecified role prey prey c-terminal ST3-DN1H required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false Fig. 7 Phosphorylation of the tyrosine-767 in the gp130 peptide was required for the interaction. The binding of the b-ST3-DN1H (lacking the first helix in the coiled coil domain) protein was not as strong as the binding of the full length protein and ST3-DN4H (320-770) did not bind at all.